The acetylene reduction assay for measuring nitrogen fixation in waterlogged soil.

Nitrogen fixation in waterlogged, soil-straw, and sandday-straw mixtures was measured with the C2H2reduction assay, the I5N-tracer technique, and the Kjeldahl method. The reduction of 6 to 15 moles C2H2 corresponded to the fixation of 1 mole N2. The theoretical ratio is 3 moles C2H2 to 1 mole N2. A ratio of 3 moles C2H2 reduced for each mole of N2 fixed was obtained when samples of sandxlay-straw were incubated under conditions that minimized effects that were due to gas diffusion through the aqueous phase. Calculations indicated that N2 at a partial pressure of 0.2 atm above the samples was not sufficient to saturate the nitrogenase enzyme of the organisms in lower layers of soil-straw samples. Thus the concentration of N2 dissolved in the aqueous phase limited nitrogen fixation. C2H2 is more soluble in water than N2; the C2H2 reduction was not as limited by the C2H2 concentration in the aqueous phase. N2 was experimentally shown to be limited at depth in a sandday-glucose system in that fixation decreased from 128 to 36 pg N/g of sandxlay incubated so that the total sample depth ranged from 0.2 to 3 cm. The C2H2 reduction assay provides a method for measuring the potential nitrogenase activity in the waterlogged soil amended with straw; however, this assay must be calibrated for specific conditions. The data also indicate that where N2 diffusion rates may limit N fixation, a normal atmosphere (80%) of N2 should be used in the experiment.